A fluorogenic, peptide-based probe for the detection of Cathepsin D in macrophages

Supplementary information The online version contains supplementary material available at https://doi.org/10.1038/s42004-023-01035-9Cathepsin D is a protease that is an effector in the immune response of macrophages, yet to date, only a limited number of probes have been developed for its detection. Herein, we report a water soluble, highly sensitive, pH insensitive fluorescent probe for the detection of Cathepsin D activity that provides a strong OFF/ON signal upon activation and with bright emission at 515 nm. The probe was synthesised using a combination of solid and solution-phase chemistries, with probe optimisation to increase its water solubility and activation kinetics by addition of a long PEG chain (5 kDa) at the C-terminus. A BODIPY fluorophore allowed detection of Cathepsin D across a wide pH range, important as the protease is active both at the low pH found in lysosomes and also in higher pH phagolysosomes, and in the cytosol. The probe was successfully used to detect Cathepsin D activity in macrophages challenged by exposure to bacteria.Engineering and Physical Sciences Research Council (EPSRC, UK) for Interdisciplinary Research Collaboration grants EP/R005257/1Medical Research Council SHIELD consortium grant MR/N02995X/

Moving into the red – a near infra-red optical probe for analysis of human neutrophil elastase in activated neutrophils and neutrophil extracellular traps

Neutrophils are the first immune cells recruited for defence against invading pathogens; however, their dysregulated activation and subsequent release of the enzyme human neutrophil elastase is associated with several, inflammation-based, diseases. Herein, we describe a FRET-based, tri-branched (one quencher, three fluorophores) near infrared probe that provides an intense OFF/ON amplified fluorescence signal for specific detection of human neutrophil elastase. The probe allowed selective detection of activated neutrophils and labelling of neutrophil extracellular traps. A tri-branched, FRET-based probe for the detection of hNE was synthesised with emission in the NIR region of the spectrum, where endogenous biomolecules have decreased absorbance, and minimal autofluorescenceUK Research & Innovation (UKRI) Engineering & Physical Sciences Research Council (EPSRC) EP/R005257/1UK Research & Innovation (UKRI) Medical Research Council UK (MRC) MR/N02995X/

A bioinspired hybrid silica–protein material with antimicrobial activity by iron uptake

A silica–protein hybrid material has been prepared by simultaneous covalent deposition of apoferritin and lactoferrin on functionalized silica. This material exhibits strong antibacterial activity against E. coli due to its high iron-uptake capacity.This work was supported by MINECO and FEDER (project CTQ2009-09344), Junta de Andalucía (project P07-FQM-02899) and BIOSEARCH SA (contract PostBIO)

Vinilsulfonas: síntesis y aplicaciones en etiquetado, inmovilización, transfección y catálisis

Tesis Univ. Granada. Departamento de Química Orgánica. Leída el 4 abril del 201

Removal of Erythromycin fromWater by Ibuprofen-Driven Pre-Organized Divinyl Sulfone Cross-Linked Dextrin

Water recycling and reuse are cornerstones of water management, which can be compromised by the presence of pollutants. Among these, pharmaceuticals can overcome standard water treatments and require sophisticated approaches to remove them. Sorption is an economically viable alternative limited by the need for sorbents with a sorption coefficient (Kd) higher than 500 L/kg. The cross-linking of dextrin (Dx) with divinyl sulfone (DVS) in the presence of 1 mmol or 5 mmol of ibuprofen (IBU) yields the insoluble polymers pDx1 and pDx5 with improved affinity for IBU and high selectivity towards erythromycin (ERY) and ERY Kd higher than 4 × 103 L/kg, when tested against a cocktail of six drugs. Characterization of the polymers shows that both pDx1 and pDx5 have similar properties, fast sorption kinetics, and ERY Kd of 13.3 × 103 for pDx1 and 6.4 × 103 for pDx5, representing 26.6 and 12.0 times the 500 L/kg threshold. The fact that new affinities and improvements in Kd can be achieved by cross-linking Dx in the presence of other molecules that promote pre-organization expands the applications of DVS cross-linked polysaccharides as sustainable, scalable, and environmentally friendly sorbents with a potential application in wastewater treatment plants (WTPs).Junta de Andalucía through the research project B-FQM-316-UGR20 funded by the Programa Operativo FEDER Andalucí

Rapid Polymer Conjugation Strategies for the Generation of pH-Responsive, Cancer Targeting, Polymeric Nanoparticles

The combination of controlled living polymerization in association with rapid and highly efficient macromolecule conjugation strategies provides a powerful tool for the synthesis of novel polymeric materials. Here functional block copolymers were rapidly and quantitatively conjugated using an efficient reaction between polymers containing a phenolic group and the 4-phenyl-3H-1,2,4-triazole-3,5(4H)-dione (PTAD) moiety and used to generate nanoparticles that encapsulated drugs. pH responsive amphiphilic block copolymers, which self-assemble into nanoparticles, were fabricated using our novel polymer conjugation strategy with the resulting system designed to promote drug release within the acidic milieu of the cancer microenvironment. The conjugation strategy also enabled the direct tagging of the nanoparticles with a range of fluorophores, targeting assets, or both with cargo release demonstrated in cancer cells.This work was funded by the European Research Council (Advanced Grant ADREEM ERC-2013-340469). W.L. ac knowledges support from the Chinese Scholarship Council. We thank the Wellcome Trust for the Multi User Equipment Grant WT104915MA

Laboratorio de Síntesis Orgánica. Guion de prácticas

Compendio de prácticas impartidas en la asignatura Laboratorio de Síntesis Orgánica, del Grado en Química

Compuestos poliméricos con propiedades inmovilizantes

Número de publicación: ES2385172 B1. Número de solicitud: 201031944.La presente invención se refiere a compuestos poliméricos con propiedades inmovilizantes sobre biomoléculas. Dichos materiales comprenden un núcleo de nanopartículas híbridas super-paramagnéticas y grupos vinilsulfona para llevar a cabo la inmovilización de las biomoléculas. Además la presente invención se refiere a un procedimiento de síntesis de dichos compuestos poliméricos.Universidad de Granad

Vinyl Sulfone Functionalization: A Feasible Approach for the Study of the Lectin−Carbohydrate Interactions

The authors acknowledge Direccion General de Investigacion Cientıfica y Tecnica (DGICYT) (CTQ2008-01754) and Junta de Andalucia (P07-FQM-02899) for financial support. A. Megia-Fernandez thanks the University of Granada for a research grant (Programa Puente).Carbohydrate-mediated molecular recognition is involved in many biological aspects such as cellular adhesion, immune response, blood coagulation, inflammation, and infection. Considering the crucial importance of such biological events in which proteins are normally involved, synthetic saccharide-based systems have emerged as powerful tools for the understanding of protein−carbohydrate interactions. As a new approach to create saccharide-based systems, a set of representative monosaccharides (D-mannose, D-glucose, N-acetyl-D-glucosamine, and L-fucose) and disaccharides (lactose, maltose, and melibiose) were derivatized at their anomeric carbon with a vinyl sulfone group spanned by an ethylthio linker. This vinyl sulfone functionalization is demonstrated to be a general strategy for the covalent linkage of a saccharide in mild conditions via Michael-type additions with the amine and thiol groups from functionalized supports and those naturally present in biomolecules. The introduction of the ethylthio linker between the biorecognizable element (i.e., saccharide) and the reactive group (i.e., vinyl sulfone) was found to preserve the functionality of the former. The capability of the vinyl sulfone saccharides for the study of lectin−carbohydrate interactions was demonstrated by (i) immobilizing them on both amine-functionalized supports (glass slides and microwell plates) and polylysine-coated glass slides to create sugar arrays that selectively bind lectins (ii) coupling to model proteins to yield neoglycoproteins that are recognized by lectins and (iii) using vinyl sulfone saccharides as tags to allow the detection of the labeled biomolecule by HRP-lectins. The above results were further put tothe test with a real case: detection of carbohydrate binding proteins present in rice (Oryza sativa)Dirección General de Investigación Científica y Técnica (DGICYT) (CTQ2008-01754)Junta de Andalucía (P07-FQM-02899)University of Granad

Non-Magnetic and Magnetic Supported Copper(I) Chelating Adsorbents as Efficient Heterogeneous Catalysts and Copper Scavengers for Click Chemistry

Novel supported chelating adsorbents bearing diverse multidentate nitrogenated ligands with strong copper(I) affinities are easily prepared in non-magnetic and magnetic variants using silica and silica-coated magnetite nanoparticles as suitable supports and the aza-Michael-type addition of vinyl sulfones as the ligation tool. These adsorbents are versatile materials with applications in the copper-catalyzed alkyne-azide cycloaddition (CuAAC) click chemistry where their complexation abilities enable them to act either as heterogeneous click catalysts when used in their complexed form or as copper(I) scavengers when used in their uncomplexed form. In the first instance, they proved to be robust and efficient heterogeneous catalysts to promote click reactions using extremely low doses and showing negligible copper leaching, particularly in the case of the silica-based non-magnetic adsorbents, allowing a simple operational protocol for their rapid and easy removal by filtration or magnetic decantation and showing good recyclability properties. In their uncomplexed form, the non-magnetic chelating adsorbents are very efficient copper scavengers that are able to remove any traces of metal contamination.Direccion General de Investigacion Cientifica y Tecnica (DGICYT) (CTQ2008-01754)Programa FPU. Junta de Andalucia (P07-FQM-02899